Abstract

Nicotinamide adenine dinucleotide (NAD+) plays a critical role in regulating numerous biological and physiological pathways including metabolism, inflammation, cancer, and aging in mammals. Here we describe a highly quantitative method with reverse-phase high-performance liquid chromatography (HPLC) for the determination of NAD+ levels in cells and tissues. This methodology provides accurate, reliable, and reproducible results of NAD+ measurement, which enables us to analyze various pathophysiological changes in NAD+ levels in vitro and in vivo.

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Acknowledgments

We thank Drs. Xuntian Jiang and Daniel Ory for mass spec analysis in the Metabolomics Facility at Washington University School of Medicine. We also thank members of the Imai lab for their critical discussions. This work was supported in part by the National Institute on Aging (AG02150), the Ellison Medical Foundation, and the Longer Life Foundation to S.I. and by institutional support from the Washington University Nutrition Obesity Research Center (P30DK056341) and the Washington University Diabetes Research and Training Center (P60DK020579). S.I. serves as a scientific advisory board member for Sirtris, a GSK company.